Generation of monospecific peptide antibodies to the DNA binding domain ofp53

The DNA binding domain (DBD) is the most mutated region of p53 in tumors an d has proven to be relatively resistant to the generation of specific antib odies. Template assembled synthetic peptide (TASP) synthesis of a peptide d erived from the DBD creates a highly immunogenic molecule without the need for large carriers such as keyhole limpet hemocyanin (KLH). In addition, a rapid means of generating monoclonal antibodies can be achieved through imm unization in conjuction with ABL/ MYC retrovirus injection into recipient m ice. In this paper, we demonstrate that an antibody generated by this means , KH2, reacts specifically with the DBD of p53. To date, this is the first example of a peptide immunogen used successfully in ABL/MYC monoclonal anti body production. KHZ is also the first example of a monospecific antibody t hat directly binds to and, by definition, assumes the conformation of the D NA binding region of p53.